Cloning and sequence analysis of a catechol 2,3-dioxygenase gene from the nitrobenzene-degrading strain Comamonas sp JS765.
Comamonas sp strain JS765 utilizes nitrobenzene as a carbon and nitrogen source. The initial attack on nitrobenzene is carried out by nitrobenzene 1,2-dioxygenase, which converts nitrobenzene to an unstable nitrohydrodiol that spontaneously decomposes to form catechol and nitrite. Catechol is then degraded via a meta cleavage pathway. We now report the cloning of a DNA fragment carrying a catechol 2,3-dioxygenase gene from JS765. Nucleotide sequence analysis revealed three open reading frames (ORFs) predicted to encode proteins of 33.6, 13.0, and 35.0 kDa. Homology searches of the deduced amino acid sequences of three proteins suggested that ORF1 encodes a LysR-type transcriptional regulator, ORF2 encodes a XylT-type ferredoxin, and ORF3 encodes a catechol 2,3-dioxygenase. The putative regulatory gene, designated cdoR, is divergently transcribed from the ferredoxin and catechol dioxygenase genes, cdoT and cdoE, respectively. The catechol 2,3-dioxygenase is most similar in amino acid sequence to the 1.2.C subfamily of extradiol dioxygenases which include 3-methylcatechol 2,3-dioxygenase from the aniline- and toluidine-degrading Pseudomonas putida UCC2, TbuE from the toluene monooxygenase pathway of Pseudomonas pickettii PKO1 and catechol 2,3-dioxygenase II from the TOL plasmid pWW15. The substrate range of the catechol 2,3-dioxygenase produced by the recombinant E. coli strains was very similar to that of the enzyme present in nitrobenzene-grown JS765, suggesting that we have cloned the catechol 2,3-dioxygenase gene required for nitrobenzene degradation.
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Department of Microbiology, College of Medicine, University of Iowa, Iowa City 52242, USA.
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001673:Biodegradation, Environmental; D050557:Catechol 2,3-Dioxygenase; D003001:Cloning, Molecular; D049308:Dioxygenases; D016946:Gram-Negative Aerobic Rods and Cocci / Q000201:enzymology*; D008969:Molecular Sequence Data; D009578:Nitrobenzenes / Q000378:metabolism*; D010105:Oxygenases / Q000235:genetics*
D009578:Nitrobenzenes; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase